As we approach the end of 2016, Shanghai Jinma has been reflecting on its progress over the past six months. This success is made possible by the support of our customers and the hard work of our employees. We are truly grateful for your trust and collaboration, which have helped us grow and achieve great results. In the first half of the year, we received numerous inquiries, and our technical team was always ready to assist with experiments and answer questions.
Our technical department has compiled a list of frequently asked questions and answers regarding the ELISA test kits (http://elisa.app17.com/shyksw-Products/T110729.html) to help you better understand the process. Below, you can find the most common questions and detailed answers:
(1) Q: Mouse acetylcholine Elisa Detection – What is the role of the blank OD value in the data processing of the kit?
A: The blank OD value is used to subtract background interference. You can either subtract it from the standard and sample values or keep it consistent throughout the experiment.
(2) Q: Why do we calculate the negative value of ACH content?
A: A negative value may occur when the sample concentration is extremely low, especially when close to the sixth point on the standard curve. This is due to the linear equation used for calculation.
(3) Q: I didn’t add one of the two blank holes. One added A liquid later, and B liquid also had stop liquid. Which one should be used?
A: For more accurate results, it's recommended to add all reagents (A, B, and stop solution) to the blank well. This helps eliminate potential interferences.
(4) Q: Is there a linear regression between standard concentration and absorbance? What could cause a low absorbance value (around 0.08)? (Rat serotonin (5-HT) Elisa Kit)
A: Yes, there should be a linear relationship. Low absorbance values could be due to sample dilution, operational errors, or instrument calibration issues. As long as they fall within the acceptable range, it’s not a problem.
(5) Q: The standard absorbance shows a linear regression, but the sample absorbance is close to the blank. No 5-HT expression in normal rat plasma. Why?
A: This might be caused by improper sample handling, dilution, or instrument settings. If using fluorescence detection, the titer could be very low.
(6) Q: What method does the Elisa kit use?
A: It uses a double-antibody sandwich method (currently temporary).
(7) Q: How much serum is needed per well?
A: Typically, 10–50 microliters per well.
(8) Q: How much unprocessed sample should be sent?
A: 100 microliters per well.
(9) Q: How should samples be stored?
A: Store fresh samples at 4°C, short-term samples at -20°C, and long-term samples at -70°C. Avoid repeated freezing and thawing.
(10) Q: What is an EP tube?
A: It's a centrifuge tube commonly used in lab settings.
(11) Q: What type of tube should be used for collecting serum?
A: Standard centrifuge tubes can be used.
(12) Q: How many samples can a 96T or 48T kit detect?
A: A 96T kit can detect up to 85 samples, while a 48T kit can detect up to 37 samples, depending on the number of standards and controls.
(13) Q: Is there any advantage to using a standard test compared to single-hole testing?
A: The standard test, also known as double-hole testing, ensures better repeatability and accuracy.
(14) Q: How to store unused ELISA plates after use?
A: If not immediately used, remove the plate from the foil bag, store in a refrigerator at 2–8°C, and avoid contamination.
(15) Q: Is an enzyme-labeled coating plate the same as an ELISA plate?
A: Yes, it is the same thing.
(16) Q: What is the general sampling amount for detecting jasmonic acid and salicylic acid in plants?
A: Around 1–3 grams of plant tissue is typically sufficient.
(17) Q: My sample exceeds the minimum detection limit. Should I dilute the standard?
A: Only if the sample concentration is extremely high. Otherwise, no need to dilute the standard unless specified in the protocol.
(18) Q: Why did my R² value drop from 0.98 to 0.93?
A: If a data point deviates significantly, it can be excluded. Keep at least five points for a reliable curve.
(19) Q: What is a blank control?
A: A blank control contains no sample, no diluent, and no reagent—just the buffer solution.
(20) Q: Is the sample diluent pre-prepared?
A: Yes, the sample diluent is already prepared.
(21) Q: Can the sample diluent be used directly to dilute serum?
A: Yes, you can add 40 μL directly into the sample.
(22) Q: Do I need to perform the test in a fume hood?
A: It’s not strictly necessary. You can perform the test anywhere as long as the lab conditions are controlled.
(23) Q: Is the washing solution concentrated or ready-to-use?
A: It is concentrated and needs to be diluted before use.
(24) Q: Can ELISA measure the titer of IgG antibodies in human platelets?
A: If ELISA is not suitable, the fluorescence method can be used instead.
(25) Q: Does an uneven ELISA plate affect the results?
A: No, the plate's surface does not impact the measurement results.
(26) Q: Can ELISA measure TNF production in cell cultures?
A: Yes, but proper grouping and calculations are required. More literature may be needed for reference.
(27) Q: What precautions should be taken when using testosterone or estriol ELISA kits?
A: Avoid adding sodium azide to horseradish-peroxidase labeled samples.
(28) Q: How should tobacco leaf samples be processed for SA detection?
A: Grind 500 mg of tobacco leaves, add 5 mL of PBS (pH 7.4), centrifuge, and collect the supernatant. Methanol is not necessary.
(29) Q: Why wasn't a standard curve made?
A: This could indicate an error in the procedure. Make sure to shake the standard solutions before use.
(30) Q: Can multiple boxes of the same ELISA kit be used without making a standard curve?
A: It's recommended to make a standard curve for each box, even if they're from the same batch. The results will be consistent.
Shanghai Jinma Experimental Equipment Co., Ltd. leverages advanced enzyme-linked immunosorbent technology, combining the expertise of domestic and international scientists. Through cutting-edge equipment and methods, we ensure quality while reducing costs. Our goal is to support researchers by saving experimental funds and ensuring reliable results, contributing to the development of science and technology in the country.
We collaborate with numerous bio-research bases and our dedicated team is committed to focusing on the production and development of ELISA kits to support your research. Let’s continue to drive innovation and discovery together!
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